Pharmacokinetics of peptide T in patients with Acquired Immunodeficiency Syndrome (AIDS)

1. The pharmacokinetics of Dalal-peptide T-NH2 (peptide T) was determined during phase I clinical trials in patients with acquired immunodeficiecy disease (AIDS) and AIDS related complex (ARC). Drug levels were determined by specific RIA, and in some cases with HPLC analysis, after intraveneous (i.v.) or intranasal (i.n.), via metered sprayer, administration.

2. The plasma kinetics appeared to be bi-phasic with a first compartment half-life of 30 to 60 minutes and a second plasma clearence rate of 4 to 6 hours, observed for both routes of administration. Peptide T, in one individual was confirmed to be present at 6 hrs in plasma, determined after HPLC isolation followed by specific RIA.

3. Bioavailabilty, determined for a 2 mg test dose in six individuals was 9.3 ± 6.9 nmol/L. Peak plasma levels of 41 ± 30 nmol/L after 10 mg i.n., 2.8 ± 5.9 nmol/L after 2mg i.n., and 0.13 ± 0.07 nmol/L after 0.4 mg i.n. were observed. In two individuals tested, peptide T was detected in CSF at levels 20% of the corresponding plasma level 90 and 145 minutes post i.v. administration. Peptide T was not detected in urine. I.N. administration was well tolerated for times up to 21 months.     

Sensitisation to Swine Leukocyte Antigens in Patients with Broadly Reactive HLA Specific Antibodies

We have previously shown that IgG HLA specific antibodies in the sera of highly sensitised patients awaiting renal transplantation can cross-react with swine leukocyte antigens (SLA). In this study we determined the frequency of patient serum IgG HLA specific antibody binding to a porcine lymphocyte panel and the likelihood of locating a cross-match negative pig donor for sensitised patients. Serum samples (n = 82) were obtained from 35 sensitised [current IgG panel reactive antibodies (PRA) > 10%] and seven nonsensitised patients awaiting renal transplantation at Addenbrooke's Hospital, Cambridge, UK. Fifty sera had IgG HLA specific PRA of 11-84%, 20 had IgG PRA of >84% and 12 had 0% PRA (negative controls). Sera were absorbed with porcine erythrocytes to remove xenoreactive natural antibodies and tested for cross-reactive IgG HLA specific antibody binding by flow cytometry against a panel of porcine lymphocytes obtained from 23 human decay accelerating factor (hDAF) transgenic pigs. A total of 1,884 cross-match combinations were tested and 369 (20%) gave a positive porcine lymphocyte cross-match. For sera from sensitised patients with IgG PRA (11-64%), only 6 of 805 (0.75%) cross-match tests were positive. In contrast, for sera from patients with high IgG PRA (>64%), 363 of 805 (45%) cross-match tests were positive (p < 0.0001). There was no difference in the frequency of positive cross-matches between patient sera with IgG PRA 65-84% and highly sensitised patient sera with IgG PRA 85-100% [156/345 (45%) vs. 207/460 (45%)]. This study demonstrates that only patient sera with broadly reactive IgG HLA specific PRA (>64%) cross-react with porcine lymphocytes. If future clinical trials of xenotransplantation are undertaken, it may be of value to select a cross-match-negative pig organ donor for such patients.     

Estimating shelf-life of drugs in solution

The use of pseudo-first-order chemical reaction kinetics and Arrhenius thermodynamic principles in estimating the effect of temperature on shelf-life of drug solutions is described. The degradation of many drugs in aqueous solutions occurs by hydrolysis. Equations using activation energy of the hydrolysis reaction (Ea) derived from Arrhenius principles, rate constants, and temperature are used to illustrate the exponential decrease in drug shelf-life proportional to an arithmetic increase in temperature. Use of the Q value, a factor based on Ea, rate constants, and temperature change, is described. The range of Ea for pharmaceutically important degradation reactions is relatively limited; thus, near room temperature, Q values of 2-5 can be used to estimate the effect of temperature change on shelf-lives of drugs in solution. To conservatively estimate effect on shelf-life when Ea is not known, a Q value of 2 is used to predict the increase in shelf-life obtained by decreasing storage temperature, and a Q of 5 is used to predict decrease in shelf-life from increasing the temperature. Sample calculations are used to show the application of equations based on these principles to practical temperature-dependent problems regarding shelf-life. Given information about shelf-life at a certain temperature, pharmacists can estimate safe shelf-life under other temperature conditions by using these equations.     

Prevalence of total hip replacement: how much demand has been met?

OBJECTIVE--To determine the prevalence of completed elective total hip replacements in a defined elderly population. DESIGN--Cross sectional postal questionnaire survey with additional data and validation from general practice and hospital records. SETTING--Six general practices in the English counties of Avon, Somerset, and Oxfordshire. SUBJECTS--A total of 7806 patients aged 65 years and over (94.7% response). RESULTS--The overall prevalence (95% confidence intervals) of elective total hip replacement was 5.3 (4.8,5.8)% Age and sex specific prevalences were 2.7 (2.0,3.5)% in men and 4.1 (3.3,4.9)% in women aged 65-74 years, and 5.2 (4.0,6.5)% in men and 8.8 (7.6,10.0)% in women aged 75 years and over. Of the 415 patients who had received elective total hip replacement, 28.2% had required bilateral surgery, 20% had received at least one operation privately, and 13% had required revision surgery. CONCLUSION--Our results show an increased level of satisfied demand for total hip replacement in elderly people compared with earlier estimates. The increasing prevalence of hip replacement is an indicator of increasing potential demand for revision procedures. Population based surveys are required to establish the level of unmet demand for primary procedures. Differences in past surgical activity may be important in interpreting the wide variation in current surgical rates.     

Gene structure and genetic localization of the PCLO gene encoding the presynaptic active zone protein Piccolo

Piccolo belongs to a family of presynaptic cytoskeletal proteins likely to be involved in the assembly and function of presynaptic active zones as sites of neurotransmitter release. Given that abnormalities in the formation of synaptic junctions are thought to contribute to cognitive dysfunction during brain development, we have analyzed and compared the gene structure of the Piccolo gene, PCLO, from humans and mice and determined their chromosomal localization. A comparison of the deduced amino acid sequence of cDNA clones encoding Piccolo from human, mouse, rat and chicken reveals the presence of distinct homology domains. Only subsets of these are also present in the structurally related active zone protein Bassoon indicating that Piccolo and Bassoon perform related but distinct functions at active zones. Characterization of the PCLO gene reveals the presence of 25 coding exons spread over 380kb of genomic DNA. The human PCLO gene maps to 7q11.23-q21.3, a region of chromosome 7 implicated as a linkage site for autism and Williams Syndrome suggesting that alterations in the expression of Piccolo or the PCLO gene could contribute to developmental disabilities and mental retardation.